Institute of Plant Science and Resources, Okayama University, Chuo 2-20-1, Kurashiki, 710-0046, Japan.
Plant J. 2019 Sep;99(6):1254-1263. doi: 10.1111/tpj.14410. Epub 2019 Jun 26.
Inter-vascular transfer in rice (Oryza sativa) nodes is required for delivering mineral elements to developing tissues, which is mediated by various transporters in the nodes. However, the effect of these transporters on distribution of mineral elements in the nodes at a cellular level is still unknown. Here, we established a protocol for bioimaging of multiple elements at a cellular level in rice node by laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS), and compared the mineral distribution profile between wild-type (WT) rice and mutants. Both relative comparison of mineral distribution normalized by endogenous C and quantitative analysis using spiked standards combined with soft ablation gave valid results. Overall, macro-nutrients such as K and Mg were accumulated more in the phloem region, while micro-nutrients such as Fe and Zn were highly accumulated at the inter-vascular tissues of the node. In mutants of nodal Zn transporter OsHMA2, Zn localization pattern in the node tissues did not differ from that of WT; however, Zn accumulation in the inter-vascular tissues was lower in uppermost node I but higher in the third upper node III compared with the WT. In contrast, Si deposition in the mutants of three nodal Si transporters Lsi2, Lsi3 and Lsi6 showed different patterns, which are consistent with the localization of these transporters. This improved LA-ICP-MS analysis combined with functional characterization of transporters will provide further insight into mineral element distribution mechanisms in rice and other plant species.
在水稻(Oryza sativa)节点中,血管间转移对于将矿物质元素输送到发育组织是必需的,这是由节点中的各种转运蛋白介导的。然而,这些转运蛋白对节点中矿物质元素在细胞水平上分布的影响尚不清楚。在这里,我们建立了一种通过激光烧蚀-电感耦合等离子体质谱(LA-ICP-MS)在水稻节点中进行细胞水平上多种元素生物成像的方案,并比较了野生型(WT)水稻和突变体之间的矿物质分布特征。通过内源 C 归一化的相对比较和使用加标标准品结合软烧蚀的定量分析,两种方法都得到了有效的结果。总体而言,大量营养元素如 K 和 Mg 在韧皮部区域积累更多,而微量元素如 Fe 和 Zn 在节点的血管间组织中高度积累。在节点 Zn 转运蛋白 OsHMA2 的突变体中,节点组织中 Zn 的定位模式与 WT 没有区别;然而,与 WT 相比,最上节点 I 中的血管间组织中 Zn 的积累较低,但第三上节点 III 中的积累较高。相比之下,三个节点 Si 转运蛋白 Lsi2、Lsi3 和 Lsi6 的突变体中的 Si 沉积呈现出不同的模式,这与这些转运蛋白的定位一致。这种改进的 LA-ICP-MS 分析结合转运蛋白的功能表征将为水稻和其他植物物种中矿物质元素分布机制提供进一步的深入了解。
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